Adenovirus infection results in a robust stimulation of the inflammatory and adaptive arms of the immune system. Initiation of the antiviral immune response depends on early recognition by sentinel immune cells such as macrophage and dendritic cells. Once engaged these cells secrete inflammatory chemokines and cytokines, they undergo maturation and migration, and they make an essential contribution to adaptive immunity through co-stimulation of T-lymphocytes. Studies from the past funding period have shown rAdV activation of APC occurs through a binary viral trigger that includes vDNA activation of a cytosolic vDNA sensor network. In addition to intrinsic viral signaling, maturation of APC also requires complementing extrinsic signals (autocrine/paracrine type I IFN and TNF1). In the current proposal we are addressing the hypothesis that cell specific vDNA signaling response networks integrate intrinsic and extrinsic signaling to generate distinct APC maturation functions. We are proposing three anchor proteins (AIM2, STING, and TBK1) are central to the intrinsic signaling cascade. Studies will define how these anchor proteins dictate the cell specific outcome in response to rAdV infection. They will also determine how the primary viral DNA sensing cascade coordinates ongoing intrinsic signaling responses with extrinsic signaling (TNF1 and type I IFNs). We are proposing that maturation of APC through these pathways defines the nature of the ensuing antiviral innate and adaptive immune responses. The knowledge gained from these studies will contribute to enhancing vaccine, gene therapy, and anticancer therapeutic applications and they will contribute to our ability to develop new treatments for viral infections and host DNA dependent autoimmune diseases.